运动对高脂膳食诱导的IR小鼠骨骼肌线粒体内膜蛋白PHB2表达和线粒体自噬的影响  

The Effect of Mitochondrial Membrane Protein PHB2 and Mitophagy on Exercise Alleviating Insulin Resistance

作　　者：齐洁[1] 牛书妍 李纯 张钧[1] QI Jie;NIU Shuyan;LI Chun;ZHANG Jun(Shanghai Normal University,Shanghai 200234,China)

机构地区：[1]上海师范大学,上海200234

出　　处：《北京体育大学学报》2022年第7期104-112,共9页Journal of Beijing Sport University

基　　金：国家自然科学基金项目“Nox4-氧化应激-自噬信号通路在运动改善胰岛素抵抗中的作用机制研究”(项目编号:31701037)。

摘　　要：目的:探讨长期规律运动对预防高脂膳食引起的IR小鼠骨骼肌线粒体中抗增殖蛋白2(PHB2)的影响。方法:6周龄雄性C57BL/6N小鼠,随机分为正常饮食安静组(ND-Con)、正常饮食运动组(ND-Ex)、高脂膳食安静组(HFD-Con)和高脂膳食运动组(HFD-Ex),每组10只。通过10周高脂饲料喂养建立IR小鼠动物模型,同时对小鼠实施中等强度跑台干预。利用IGTT和IITT试验评价IR动物模型及运动预防IR的效果。通过DHE染色观察骨骼肌ROS水平,DCFH-DA试剂盒检测骨骼肌线粒体ROS生成速率,试剂盒检测骨骼肌线粒体CAT和T-AOC水平,western blot检测骨骼肌线粒体自噬相关基因PHB2、PINK1、Parkin、LC3蛋白表达水平。结果:1)与ND-Con组比较,IGTT和IITT试验结果表明HFD-Con组血糖曲线下面积(AUC)显著增加(P<0.01);与HFD-Con组相比,HFD-Ex组AUC显著下降(P<0.01,P<0.05)。2)与ND-Con组比较,HFDCon组骨骼肌ROS水平和线粒体ROS生成速率均显著增加(P<0.01),骨骼肌线粒体CAT、T-AOC水平均显著下降(P<0.01);与HFD-Con组比较,HFD-Ex组骨骼肌及线粒体氧化应激指标均显著改善(P<0.01)。3)与ND-Con组比较,HFD-Con组骨骼肌线粒体相关基因PHB2、PINK1、Parkin蛋白表达量以及LC3Ⅱ/LC3Ⅰ比值均显著下降(P<0.01);与HFD-Con组比较,HFD-Ex组骨骼肌线粒体自噬相关蛋白表达水平均显著增加(P<0.01)。结论:10周跑台运动可抑制由高脂膳食引起的骨骼肌线粒体过度氧化应激,并上调骨骼肌线粒体自噬标志物PHB2表达水平,同时增强线粒体自噬途径PINK1/Parkin信号转导,预防高脂膳食诱发小鼠形成胰岛素抵抗。This study aims to investigate the effects of prohibitin 2(PHB2)on improving insulin resistance in high-fat-diet mice through treadmill exercise.Methods:Forty C57BL/6N mice were randomly assigned to 4 groups:1)sedentary rats fed with normal diet group(ND-Con group);2)normal diet mice treadmill for 60 min/time(ND-Ex group);3)sedentary rats fed with high-fat diet group(HFD-Con group);4)high-fat diet mice treadmill for60 min/time(HFD-Ex group).The HFD-Con group was fed with high-fat diet for 10 weeks to induce insulin resistance(IR).The animal model was evaluated by IGTT and IITT.The effects of exercise on skeletal muscle ROS,oxidative stress level of mitochondria(ROS,CAT and T-AOC),PHB2、PINK1、Parkin protein levels in mice skeletal muscle mitochondria were observed to explore the mechanism of exercise improving IR.Results:1)after 10-week high-fat diet,glucose utilization and insulin tolerance in the HFD-Con group significantly decreased(P<0.01),which indicated that the animal model of IR was established successfully.Compared with the HFD-Con group,the glucose utilization and insulin tolerance in the HFD-Ex group obviously improved after 10 weeks(P<0.01).2)Compared with the ND-Con group,in the HFD-Con group,the ROS level in skeletal muscle and mitochondria markedly increased(P<0.01),and the CAT and T-AOC activity in skeletal muscle mitochondria markedly decreased(P<0.01).When compared with the HFD-Con group,the ROS in skeletal muscle and oxide stress in skeletal muscle mitochondria markedly improved in the HFD-Ex group(P<0.01).3)Compared with the ND-Con group,PHB2,PINK1,Parkin and LC3Ⅱprotein levels were dramatically down-regulated(P<0.01).When compared with the HFD-Con group,the mitophagy related genes detected in skeletal muscle mitochondria were markedly up-regulated in the HFD-Ex group(P<0.01).Conclusion:10-week treadmill training can alleviate the damage degree of skeletal muscle cells by inhabiting oxidative stress,which can up-regulate PHB2 level and enhance the signal transduction of PINK1/Parkin in ske

关 键 词：长期规律运动 骨骼肌 线粒体胰岛素抵抗 活性氧 抗增殖蛋白2 PINK1/Parkin信号通路 

分 类 号：G804.2[文化科学—运动人体科学]